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Acta Biologica Hungarica

, Volume 65, Issue 4, pp 405–413 | Cite as

Detection of DNA Damage Caused by Cryopreservation using a Modified SCGE in Large Yellow Croaker, Pseudosciaena crocea

  • Xiang-Rong Xu
  • Fu-Qing Tan
  • Jun-Quan Zhu
  • Ting Ye
  • Chun-Lin Wang
  • Yi-Feng Zhu
  • Hans-Uwe Dahms
  • Fan Jin
  • Wan-Xi YangEmail author
Open Access
Article

Abstract

We used single-cell gel electrophoresis (SCGE) to detect the integrity of sperm DNA of the teleost large yellow croaker, Pseudosciaena crocea, cryopreserved with Cortland solution and a range of 5% to 30% DMSO concentrations in order to test how sperm cryopreservation affected the DNA stability of nuclei. Electrophoresis was conducted for 60 min at 130 mA and 15 V. The comet images were analyzed with software CometScore 1.5, and parameters such as comet length, tail length and percentage DNA in the tail were obtained. Then the comet rate and damage coefficient were calculated. Results demonstrated that there were no significant differences in motility, comet rate and damage coefficient between fresh sperm and cryopreserved sperm stored in 5%, 10%, 15% and 20% DMSO, while the sperm cryopreserved with 25% and 30% DMSO had a lower motility, higher comet length and damage coefficients than those of fresh sperm. There was a positive correlation between comet rate of cryopreserved sperm and the concentration of DMSO. Our results demonstrate that toxicity of the cryoprotectant is the main cause of DNA damage in cryopreserved sperm nuclei.

Keywords

Pseudosciaena crocea sperm cryopreservation genetic damage single-cell gel electrophoresis 

Notes

Acknowledgements

We thank the following projects for their support: K. C. Wong Magna Fund in Ningbo University, The Scientific and Technical Project of Zhejiang Province, National Natural Science Foundation of China (Grant Nos. 41276151, 31272642 and 31072198).

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© Akadémiai Kiadó, Budapest 2014

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • Xiang-Rong Xu
    • 1
  • Fu-Qing Tan
    • 2
  • Jun-Quan Zhu
    • 3
  • Ting Ye
    • 3
  • Chun-Lin Wang
    • 3
  • Yi-Feng Zhu
    • 3
  • Hans-Uwe Dahms
    • 4
  • Fan Jin
    • 1
  • Wan-Xi Yang
    • 5
    Email author
  1. 1.Department of Reproductive Endocrinology, Key Laboratory of Reproductive Genetics, Women’s HospitalZhejiang University School of MedicineHangzhouChina
  2. 2.The First Affiliated Hospital, College of MedicineZhejiang UniversityHangzhouChina
  3. 3.Key Laboratory of Applied Marine Biotechnology by the Ministry of EducationNingbo UniversityNingboChina
  4. 4.Dept. of Biomedical Science and Environmental BiologyKMU - Kaohsiung Medical UniversityKaohsiungTaiwan, Republic of China
  5. 5.The Sperm Laboratory, College of Life ScienceZhejiang UniversityHangzhouChina

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